Hepatitis C virus (HCV) is a single-stranded RNA virus of roughly 9.4 kb estimated to affect 200 million people worldwide. In the U.S., where it is estimated that 4.1 million are infected, HCV accounts for 60-70% of chronic hepatitis and 50% of cirrhosis, end-stage liver disease, and hepatocellular carcinoma, causing an estimated 12,000 deaths annually. From 1994 to 2007, annual hospitalizations for HCV rose 10.3-fold, while total healthcare spending increased from $627 million to $6.9 billion in HCV mono-infected patients and from $63 million to $655 million in HIV/HCV co-infected patients. Approximately 75% of HCV-infected people in the U.S. are unaware of their condition and are at increased risk for late-stage complications of cirrhosis and liver cancer. Patient access outside of institutional healthcare systems is limited by the high cost of testing and treatment, particularly for the uninsured and underinsured - only 54% of HCV treatment candidates have any type of insurance coverage. Patients at Veterans Administration hospitals, urban Native American populations, and ethnic minorities in the U.S. have disproportionately high rates of chronic HCV infection and complications. Current standard-of-care combination antiviral therapy (ribavirin/peg-interferon) eradicates HCV in many patients, while newly FDA-approved HCV-specific therapies greatly increase the likelihood and speed of positive treatment outcomes. Prerequisites for successful administration of current and emerging therapeutic regimens are genotyping and measurement of viral loads, both of which help determine treatment course and duration. Determination of baseline HCV RNA levels is critical for monitoring patient response to therapy, and recent evidence suggests that personalizing treatment based on initial viral load and virological response increases success rates. As the HCV standard of care evolves, identifying people infected with HCV takes on even greater importance, including a renewed push for screening programs to identify and treat undiagnosed patients, as well as for rapid point of care testing. This proposal develops the EOSCAPE-CLEF (Chelated Lanthanide Emission Fingerprinting), a low-cost, point of care (POC) alternative to conventional HCV RNA testing, combining quantitative detection by signal amplification with sensitive and specific genotyping using a pioneering signal processing algorithm based on the characteristic light emission profiles of individual chelate-lanthanide pairs conjugated to a stringent molecular beacon-style probe. The enclosed-cartridge format system is anticipated to reduce costs from a combined $400 for quantitation and genotyping to less than $35 while also reducing the turnaround time to 30 minutes. The assay runs in a single-use enclosed cartridge based on the Applicant Organization's monolithic slit capillary array fluidic microactuator (mSCAFA) technology.